title: An HLA-modified ovarian cancer cell line induced CTL responses specific to an epitope derived from claudin-1 presented by HLA-A*24:02 molecules.
abstract: In an attempt to induce cytotoxic T lymphocytes (CTLs) that react to ovarian cancer cells, we isolated a CTL clone that specifically recognizes claudin-1 in an HLA-A*24:02-restricted manner. Naïve CD8(+) T lymphocytes were obtained from a healthy adult donor and stimulated twice in vitro with HLA-modified TOV21G cells that were originally derived from an ovarian clear-cell carcinoma line. The TOV21G modification involved RNAi-mediated gene silencing of intrinsic HLA molecules and lentiviral transduction of a synonymously mutated HLA-A*24:02. Then, cDNA library construction using mRNA extracted from the parental TOV21G cells and subsequent expression cloning were conducted. These experiments revealed that a CTL clone obtained from the bulk culture recognized a minimal epitope peptide RYEFGQALF, which was derived from an autoantigen claudin-1 presented by HLA-A*24:02 molecules. This clone exhibited cytolytic activities against three ovarian cancer cell lines and normal bronchial epithelial cells in an HLA-A*24:02-restricted manner. Our data indicate that HLA-modified cancer cells can be used as an artificial antigen-presenting cell to generate antigen-specific CTLs in a manner restricted by an HLA allele of interest.
authors.affiliation: Division of Immunology, Aichi Cancer Center Research Institute, Nagoya 464-8681, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Kondo
authors.forename: Ayako
authors.initials: A
authors.lastname: Demachi-Okamura
authors.forename: Tomoya
authors.initials: T
authors.lastname: Hirosawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Maki
authors.forename: Mitsugu
authors.initials: M
authors.lastname: Fujita
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Uemura
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Akatsuka
authors.forename: Eiko
authors.initials: E
authors.lastname: Yamamoto
authors.forename: Kiyosumi
authors.initials: K
authors.lastname: Shibata
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Ino
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Kikkawa
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Kuzushima
mesh_terms: D018262:Adenocarcinoma, Clear Cell / Q000276:immunology*; D000328:Adult; D045744:Cell Line, Tumor; D062445:Claudin-1 / Q000276:immunology / Q000378:metabolism*; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism*; D005260:Female; D059871:HLA-A24 Antigen / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008213:Lymphocyte Activation; D010051:Ovarian Neoplasms / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D019076:Transgenes / Q000235:genetics
chemical_list: D062445:Claudin-1; D018984:Epitopes, T-Lymphocyte; C438506:HLA-A*24:02 antigen; D059871:HLA-A24 Antigen; D010446:Peptide Fragments
keywords: A24-293T; B-LCL; EBV-transformed B lymphoblastoid cell line; HLA-A24 expressing HEK-293T; NHBE; aAPC; artificial antigen-presenting cell; normal human bronchial epithelial